The Diagnostic Value of LINC00426 in Type 2 Diabetes and Diabetic Kidney Disease and its Regulatory Effects on Renal Cells

Summary

Downregulation of LINC00426 promotes the progression of diabetic kidney disease (DKD) by inducing apoptosis in renal cells and increasing inflammation.

Abstract

The study investigates the diagnostic value of LINC00426 in type 2 diabetes (T2DM) and diabetic kidney disease (DKD), along with its regulatory effects on renal cells. The study included 280 T2DM patients (146 cases of DKD), and 186 healthy controls who visited during the same period. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) confirmed that serum LINC00426 is downregulated in patients with T2DM and DKD. Receiver operating characteristic (ROC) analysis assessed LINC00426’s diagnostic value for T2DM and DKD. Multivariate logistic regression identified independent DKD risk factors. LINC00426 showed diagnostic value for both diseases and negatively correlated with the urine albumin-to-creatinine ratio (UACR). Its downregulation independently predicted DKD development. A high-glucose (HG) environment inhibits LINC00426 expression in HK-2 and HGMC cells. CCK8 assay, flow cytometry, and enzyme-linked immunosorbent assays confirmed that HG also suppressed proliferation while promoting apoptosis and inflammation. In contrast, overexpression of LINC00426 enhances the proliferative capacity of HK-2 and HGMC cells, inhibits apoptosis, and reduces inflammatory responses. Downregulation of LINC00426 may exacerbate DKD progression by inhibiting the proliferation of renal cells (HK-2 and HGMC) and promoting apoptosis and inflammatory responses.

Introduction

Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder¹. Reports indicate that T2DM and its complications have become a significant global public health concern^2,3, posing threats to human health and societal development. Due to unhealthy lifestyle habits acquired over time⁴, the prevalence of T2DM has been increasing annually⁵. Diabetic nephropathy (DN) was renamed diabetic kidney disease (DKD) in 2007^6,7. As a common complication of T2DM⁸, its inc....

Protocol

This study has been approved by the Medical Ethics Committee of The Third Xiangya Hospital of Central South University. All patients and their families expressed support and signed written informed consent forms.

Sample Collection
The sample size was calculated using G*Power software. The significance level (α) was set at 0.05, statistical power (1-β) at 0.8, effect size (f) at 0.25, and a 10% dropout rate was considered. The calculation results indicated that at least 123 participants per group were required when studying the control group, T2DM, and DKD populations. In the actual study, 186 participants we....

Results

Comparison of Baseline Characteristics Between Healthy Individuals and T2DM Patients
The study enrolled 280 T2DM patients and 186 healthy individuals as controls. Analysis of the participants' medical records (see Table 1) revealed that the mean age of the control group was 58.91 ± 17.63 years, compared to 59.50 ± 17.14 years for the T2DM patients. However, there were no significant differences in gender or obesity between the two groups of patients. Further comparison revealed that,.......

Discussion

DKD represents a severe microvascular complication in T2DM. Its earliest manifestation is the presence of microalbuminuria; detecting this in the early stages could potentially delay DKD progression. However, microalbuminuria lacks sensitivity in routine urine analysis and is consequently frequently overlooked⁴³. This study found that LINC00426 is downregulated in T2DM patients and possesses diagnostic value for both T2DM and DKD. Furthermore, LINC00426 may prevent DKD by reducing apoptosis in HG .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.25% trypsin	Gibco	25200056
Annexin V-FITC Apoptosis Detection Kit	Beyotime	C1062S
Antibiotics for cell culture	Gibco	R-015-10
Aristo specific protein detector	GOLDSITE	GPP-100
Bax Antibody	Cell signaling Technology	2772
BCA protein assay kit	Beyotime	P0013B
Bcl-2 (124) Mouse Monoclonal Antibody	Cell signaling Technology	15071
beta-Actin (13E5) Rabbit Monoclonal Antibody	Cell signaling Technology	4970
Blood glucose monitor	Yuwell	710
Caspase-3 Antibody	Cell signaling Technology	9662
CCK-8 reagent	Sigma	96992
ChamQ Universal SYBR qPCR Master Mix	Vazyme	Q711
Chloroform	Sigma-Aldrich	C2432
Clarity Western ECL Substrate	Bio-Rad	1705061
Constant-temperature cell incubator	SaiLun	DHP-9032
D-Glucose	MedChemExpress	HY-B0389
Flow cytometer	Beckman coulted	CytoFLEX	Chloroform
Fluorescent quantitative PCR instrument	Roche	Light Cycler 96
Foetal bovine serum	Sigma-Aldrich	F0193
Fully automated glycated haemoglobin analyser	Hanyu medical	HLC-723G11	Phenol
Gene PCR amplifier	Bio Rad	S1000
Glomerular mesangial cells, HGMC	Sunncell Biotechnology Co., Ltd.	SNP-H252
Human Basic IL-1 beta ELISA Kit	Invitrogen	ECH002
Human IL-6 ELISA Kit	Invitrogen	EH2IL6
Human kidney epithelial cells, HK-2	Sunncell Biotechnology Co., Ltd.	SNL-165
Human TNF alpha ELISA Kit	Invitrogen	KAC1751
IkappaB alpha (L35A5) Mouse Monoclonal Antibody	Cell signaling Technology	4814
K-SFM medium	Invitrogen	17005-042
Lipofectamine 3000	Invitrogen	L3000015
Mannitol	Sigma-Aldrich	M9647
Multifunctional enzyme-linked immunosorbent assay reader	TECAN	GENios Plus
NF-kappaB p65 (D14E12) Rabbit Monoclonal Antibody	Cell signaling Technology	8242
non-fat milk	Biosharp	BS102-500g
Opti-MEM™ I Reduced Serum Medium	Invitrogen	31985062
P3000 Reagent	Invitrogen	L3000-015
PBS buffer	Sangon Biotech	E607080
Phenol	Sigma-Aldrich	P1037
Phospho-NF-kappaB p65 (Ser536) (93H1) Rabbit Monoclonal Antibody	Cell signaling Technology	3033
Primary cell complete medium	Sunncell Biotechnology Co., Ltd.	SNPM-H252
Prime Script RT reagent Kit	Takara	RR037A
PVDF membranes	Merck Millipore	SE1M003M00
rotease and phosphatase inhibitor cocktail	Beyotime	P1045
Trizol	Invitrogen	15596026CN
Ultraviolet spectrophotometer	Thermo	Nanodrop 2000